Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography–mass spectrometry

Luo, Baozhu; Groenke, Karsten; Takors, Ralf; Wandrey, Christian; Oldiges, Marco

doi:10.1016/j.chroma.2007.02.034

Cited by 434 publications

(419 citation statements)

References 55 publications

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“…Presently mass spectrometry (MS) has become more popular as the detection method, thereby coupled to GC, capillary electrophoresis (CE) or HPLC as initial separation step. More advanced LC-MS/MS methods allow more accurate, robust and sensitive analyses Luo et al, 2007). For application of LC-MS/MS especially the rigorous use of stable isotope (e.g.…”

mentioning

confidence: 99%

Development and application of a differential method for reliable metabolome analysis in Escherichia coli

Taymaz-Nikerel

Mey

Ras

et al. 2009

Analytical Biochemistry

145

129

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confidence: 99%

Development and application of a differential method for reliable metabolome analysis in Escherichia coli

Taymaz-Nikerel

Mey

Ras

et al. 2009

Analytical Biochemistry

145

129

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“…We are particularly interested in analyzing such compounds because plant cell wall biosynthesis involves a range of phosphorylated precursors, mainly NDPsugars and hexose-Ps (Feingold and Barber, 1990;Fry 2000Fry , 2004Seifert, 2004;Somerville et al, 2004;Sharples and Fry, 2007). For example, Huck et al (2003) and Luo et al (2007) were able to separate, respectively, six and 28 intracellular metabolites involved in glycolysis, the oxidative pentose-P pathway, and the tricarboxylic acid cycle, some of which are phosphorylated. Bajad et al (2006) were able to separate a large number of water-soluble cellular metabolites by hydrophilic interaction chromatography, but this method does not appear to separate isomers.…”

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confidence: 99%

Quantifying the Labeling and the Levels of Plant Cell Wall Precursors Using Ion Chromatography Tandem Mass Spectrometry

et al. 2010

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The biosynthesis of cell wall polymers involves enormous fluxes through central metabolism that are not fully delineated and whose regulation is poorly understood. We have established and validated a liquid chromatography tandem mass spectrometry method using multiple reaction monitoring mode to separate and quantify the levels of plant cell wall precursors. Target analytes were identified by their parent/daughter ions and retention times. The method allows the quantification of precursors at low picomole quantities with linear responses up to the nanomole quantity range. When applying the technique to Arabidopsis (Arabidopsis thaliana) T87 cell cultures, 16 hexose-phosphates (hexose-Ps) and nucleotide-sugars (NDP-sugars) involved in cell wall biosynthesis were separately quantified. Using hexose-P and NDPsugar standards, we have shown that hot water extraction allows good recovery of the target metabolites (over 86%). This method is applicable to quantifying the levels of hexose-Ps and NDP-sugars in different plant tissues, such as Arabidopsis T87 cells in culture and fenugreek (Trigonella foenum-graecum) endosperm tissue, showing higher levels of galacto-mannan precursors in fenugreek endosperm. In Arabidopsis cells incubated with [U-13 C Fru ]sucrose, the method was used to track the labeling pattern in cell wall precursors. As the fragmentation of hexose-Ps and NDP-sugars results in high yields of [PO 3 ] 2 and/or [H 2 PO 4 ] 2 ions, mass isotopomers can be quantified directly from the intensity of selected tandem mass spectrometry transitions. The ability to directly measure 13 C labeling in cell wall precursors makes possible metabolic flux analysis of cell wall biosynthesis based on dynamic labeling experiments.Plant cell walls are the most abundant renewable resources (Pauly and Keegstra, 2008a). Much of the current biotechnological research on plant cell wall synthesis involves manipulating these biosynthetic processes to obtain higher concentrations of starches or oil, which show much promise in biofuel production, or to alter cell wall composition for easier breakdown. A detailed knowledge of these processes is essential to understanding and utilizing plant cell wall materials as well as for progress in understanding plant growth and structural development (Pauly and Keegstra, 2008b). However, research into cell wall biosynthesis has been hindered by our limited understanding of the metabolic processes that produce cell walls and particularly their regulation. Progress in this area is limited by the difficulty of differentiating among the compounds involved and of analyzing the fluxes through the biochemical network of wall biosynthesis. Many of the metabolic steps involve isomeric sugars, including hexose-Ps and nucleotidesugars (NDP-sugars) that serve as direct precursors to plant cell wall biosynthesis. Separate quantification of these sugars has been difficult to achieve.Much of the current research on identifying and differentiating among different metabolic pathways involves the use of chromato...

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“…Tandem mass spectrometry (MS/MS) and exact mass measurements of intracellular metabolites have led to the identification of metabolites for improved understanding of biological pathways and discovery of potential biomarkers [17][18][19][20][21]. MS/MS involving collision-activated dissociation (CAD) has been used to obtain structural information about phosphate-containing metabolites [17][18][19][20][22][23][24]. In such experiments, negative-mode ionization is generally preferred due to the acidic character of phosphate groups [17][18][19][20][22][23][24][25].…”

mentioning

confidence: 99%

“…MS/MS involving collision-activated dissociation (CAD) has been used to obtain structural information about phosphate-containing metabolites [17][18][19][20][22][23][24]. In such experiments, negative-mode ionization is generally preferred due to the acidic character of phosphate groups [17][18][19][20][22][23][24][25]. Negative-mode CAD of phosphate-containing metabolites often results in limited fragmentation, possibly due to the absence of a mobile proton [26], and thereby provides limited structural information because it mainly results in cleavages at phosphate groups, or charge-directed fragmentation [18,19,23].…”

mentioning

confidence: 99%

“…In such experiments, negative-mode ionization is generally preferred due to the acidic character of phosphate groups [17][18][19][20][22][23][24][25]. Negative-mode CAD of phosphate-containing metabolites often results in limited fragmentation, possibly due to the absence of a mobile proton [26], and thereby provides limited structural information because it mainly results in cleavages at phosphate groups, or charge-directed fragmentation [18,19,23]. However, Gross and coworkers have recently shown that highenergy CAD of metabolite anions in a matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometer allows differentiation between isomeric metabolites such as adenosine 5=-triphosphate (ATP) and 2=-deoxyguanosine 5=-triphosphate (dGTP) [25].…”

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confidence: 99%

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Characterization of phosphate-containing metabolites by calcium adduction and electron capture dissociation

Liu

Yoo

Håkansson

2008

J. Am. Soc. Mass Spectrom.

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Several phosphate-containing metabolites, including nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), adenosine 5=-diphosphate ribose (ADP-r), adenosine 5=-triphosphate (ATP), and guanosine 5=-triphosphate (GTP), have been characterized with electron capture dissociation (ECD) and sustained off-resonance irradiation collision-activated dissociation (SORI-CAD) tandem mass spectrometry (MS/MS) in positive-ion mode. Calcium complexation was used to successfully produce abundant doubly charged cationic precursor ions with or without hydration. This approach enabled application of ECD to acidic metabolites for the first time. Fragmentation pathways observed in ECD and SORI-CAD of calcium-adducted phosphate-containing metabolites were complementary. Unique fragmentation was observed in ECD compared to SORI-CAD MS/MS, including ribose cross-ring cleavage for NAD and NADP, and generation of hydrated product ions, including cross-ring fragments, for hydrated ATP and GTP. A combination of ECD and CAD appears promising for maximizing structural information about metabolites. Due to the wide range of metabolites in a metabolic network, and their chemical diversity, it is highly challenging to establish analytical tools for identifying and quantifying all of them. Although nuclear magnetic resonance, Fourier transform infrared, and Raman spectroscopies are widely used for metabolite analysis [5, 9 -13], mass spectrometry (MS) has emerged as a more suitable technology for measurement of metabolites because of its wide dynamic range (10 4 -10 6 ), good sensitivity (nM), and ability to detect a diverse number of molecular species [14 -16].Phosphate-containing metabolites are mainly involved in the central carbon metabolism, which consists of glycolysis, the pentose-phosphate pathway, the tricarboxylic acid cycle, and their corresponding cofactors. Tandem mass spectrometry (MS/MS) and exact mass measurements of intracellular metabolites have led to the identification of metabolites for improved understanding of biological pathways and discovery of potential biomarkers [17][18][19][20][21]. MS/MS involving collision-activated dissociation (CAD) has been used to obtain structural information about phosphate-containing metabolites [17][18][19][20][22][23][24]. In such experiments, negative-mode ionization is generally preferred due to the acidic character of phosphate groups [17][18][19][20][22][23][24][25]. Negative-mode CAD of phosphate-containing metabolites often results in limited fragmentation, possibly due to the absence of a mobile proton [26], and thereby provides limited structural information because it mainly results in cleavages at phosphate groups, or charge-directed fragmentation [18,19,23]. However, Gross and coworkers have recently shown that highenergy CAD of metabolite anions in a matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometer allows differentiation between isomeric metabolites such as adenosine 5=-triphosphate (ATP) and 2=-deoxyguano...

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Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography–mass spectrometry

Cited by 434 publications

References 55 publications

Development and application of a differential method for reliable metabolome analysis in Escherichia coli

Development and application of a differential method for reliable metabolome analysis in Escherichia coli

Quantifying the Labeling and the Levels of Plant Cell Wall Precursors Using Ion Chromatography Tandem Mass Spectrometry

Characterization of phosphate-containing metabolites by calcium adduction and electron capture dissociation

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